Method and apparatus for automated rapid immunohistochemistry

ABSTRACT

A method of biochemical processing may, in certain embodiments, involve the steps of use of a consumable biochemical process element and a consumable biochemical process alterable information memory element associated therewith, such as a substance withdrawal consumable biochemical process alterable information memory element, that may be queried and at least some of whose information may be changed as a result of actions conducted during processing of a biochemical test sequence. Accordingly, advantages relative to knowing how much of a consumable biochemical process element has been used, and how much may be available during a biochemical test sequence, may be achieved.

This application is the United States National Stage of InternationalApplication No. PCT/US2006/015023, filed Apr. 21, 2006, published as WO2006/116039 on 2 Nov. 2006, and claiming the benefit of U.S. ProvisionalPatent Application No. 60/673,486, filed Apr. 21, 2005, each herebyincorporated by reference herein.

1. Technical Field

This invention relates to the field of automated sample testing such asmay be used in biochemistry, perhaps including cytochemistry,histochemistry, and the like. Specifically, it relates to systems anddevices that have particular mechanical arrangements and attributes.Such systems and devices may be particularly appropriate for use in asurgical or operative environment, where rapids results may benecessary. Furthermore, this application addresses only certain aspectsof the technology disclosed. Other aspects are addressed in theconcurrently filed applications entitled: “Enhanced Fluidic Method andApparatus for Automated Rapid Immunohistochemistry” filed this same dayand accorded serial number PCT/US2006/015020, “Parallel ProcessingFluidic Method and Apparatus for Automated Rapid Immunohistochemistry”filed this same day and accorded serial number PCT/US2006/015269, and“Wicking Cassette Method and Apparatus for Automated RapidImmunohistochemistry” filed this same day and accorded serial numberPCT/US06/15017. Each of these are hereby incorporated by reference aswell as the priority filing (which this filing claims the benefit of),US Provisional Application No. 60/673,486 entitled “Method and Apparatusfor Automated Rapid Immunohistochemistry”.

2. Background

Frequently during surgery, tissue biopsy samples may be removed from apatient and sent from the operating room to a pathology laboratory foranalysis, for example by frozen tissue section diagnosis. In addition,methodology for frozen tissue section diagnosis may consist of freezingtissue in a pathology lab, sectioning the frozen tissue, and performingstandard Hematoxylin and Eosin (H&E) staining. H&E may be ageneral-purpose stain for helping a medical pathologist diagnose tissuepathologies. However, H&E staining may have a number of limitations, forexample that it may be a non-specific tissue stain, and may not identifyspecific proteins in tissue. Such identification of specific proteins intissue, for example by using a procedure sometimes referred to asimmunohistochemistry (IHC), may help a pathologist diagnose numerousintraoperative tissue pathologies. Examples may include sentinel lymphnode biopsies (for potential metastatic carcinomas and melanomas),undifferentiated tumors (potential carcinomas, lymphomas, andmelanomas), and biopsies of margins (looking at the edges of excisedtissue to see if the entire tumor has been removed).

A problem may be that current automated IHC may require 60 to 120minutes, which may be too long to be useful during intraoperativeprocedures. Intraoperative guidelines, such as those provided by theCollege of American Pathologists, may typically recommend reportingpathology data to the surgeon within approximately 20 minutes. Anotherconcern is the economics of manufacture, as well as the ease of use.Operators can sometimes make mistakes and so a simplified manner ofoperation is desired.

It often may be difficult to examine unstained cell and tissuepreparations with a microscope, for example perhaps due to a lack ofcontrast between individual cells and the background matrix, or perhapsbetween individual parts of cells. To improve such contrast, researchersmay apply stains to cell and tissue specimens to be examined. Suchstains may be adsorbed differently by various structures in cells,perhaps such that the contrast between the different cell structures maybe improved.

Staining tissue specimens may be a nontrivial, time-consuming process.Often, a number of different staining and rinsing stages may berequired. Each stage may require a specific amount and perhaps differenttypes of reagent or buffer and may take a specific amount of time.Furthermore, at the completion of a test sequence removed material canbe a hazardous material that requires specific handling and disposal.Thus, trained technicians often may be employed to perform suchoperations. Furthermore, hospitals and laboratories may be required tostain large numbers of tissue specimens with different reagents orsubstances. Thus, it may be desirable to automate the tissue specimenstaining process and to make insertion of desired substances easier andmore foolproof. By automating the process, expensive human labor may beeliminated and the probability of an error occurring during the stainingprocess may be reduced. Accordingly, some manufacturers have introducedequipment for the automated staining of tissue specimens on microscopeslides.

However, existing automatic staining devices may not be simple to useand their internal workings in the complex. Such existing automaticstaining devices may required arcane programming commands andcomplicated procedures, which may require extensive user training beforesuch devices can be operated effectively. Waste material can alsorequire special handling. It therefore may be desirable to simplify theoperation of an automatic staining device as well as its manufacture.

As mentioned earlier, though, existing automatic staining devices cantake a significant amount of time to achieve a desired result. Whenusing interacting or perhaps binding substances, such as antibodies, ormore generally reagents, the substance used that may take a significantperiod of time to achieve its chemical result relative to anintraoperative procedure. For example, a typical reagent binding profileusing an accelerated incubation period can take in excess of 60 minutesor the like. This is usually too long to leave a patient exposed and soit is not uncommon for the patient to be sewn back up and asked toreturn once results are available. Furthermore it may not be practicalto run a bulk processing system for only one or two samples. While thistesting time period may be necessary in order to achieve an amount ofbinding or other interaction desired with most substances, such a periodof time is not typically acceptable from the perspective of performingan intraoperative procedure on a patient. Beyond merely the chemicalinteraction time period, the entire process can take even significantlylonger. Thus it is not uncommon for many staining or other biochemicalprocedures to require at least one hour in order to yield the desiredresults.

Furthermore, the entire process may be fairly involved. For example, abiochemical process can sometimes involve steps including: subjecting asample to a first antibody substance, perhaps driving the antibodysubstance around with an air knife to blow air across the surface of thesample, withdrawing the antibody substance, rinsing the sample with abuffer, subjecting the sample to a second antibody substance, perhapsagain driving the antibody substance around with an air knife,withdrawing the second antibody substance, again rinsing the sample witha buffer, subjecting the sample to a chromogen substance, withdrawingthe first chromogen substance, again rinsing the sample with a buffer,withdrawing the second chromogen substance, subjecting the sample to acounterstain, withdrawing it counterstain, and then perhaps againrinsing the sample with a buffer. Each of these steps may take asignificant amount of time in and of themselves, and may result in thesum of the entire procedure taking an inordinate amount of time. Infact, it may not be uncommon for such involved procedures to take 90minutes or more. Although there may have been efforts to shorten thistime period, the simple fact of the chemistry involved may have focusedthese efforts to some degree on speeding up the mechanical processesinvolved.

One process which may be known to speed up the chemical process,however, is to heat a sample and the substance applied. In this type ofa system, a reagent may be heated and this may reduce the reagent-tissueinteraction period. Disadvantages to heating may include the fact thatmany reagents and some samples may not react well to heating.

While the use of an air knife to blow air across the surface of areagent and to drive the reagent or other substance around on thesurface of the sample may have accomplished some shortening of theoverall process, it remains a fact that even when this function isemployed, the procedures still require long time periods on the order to60 to 120 minutes. Thus, one of the challenges and one of thelimitations of many of the automated histochemical and other suchsystems previously in use is the fact that they simply do not yieldtheir results in a short enough time period in order to provide systemsthat can be used effectively in an intraoperative environment. Prior tothe present invention, it may have even been perceived as a necessaryincident to the basic chemistry that such tests required this long atime frame. In view of the foregoing, there is a need for theavailability of an automated rapid IHC or other such system that wouldallow IHC or the like to be performed within 20 minutes or less.Automated rapid IHC or other such biochemical tests are, of course, alsodesired by research laboratories for frozen tissues and the like.

3. SUMMARY DISCLOSURE OF THE INVENTION

In embodiments, the present invention involves a self contained rapidsample processing system such as shown in FIG. 1. This system can beeconomical to manufacture and may be easily used within an operatorenvironment. Embodiments can overcome problems that have seemedinsurmountable perhaps by approaching the problem from a very differentperspective. The present invention presents systems in a variety ofembodiments through which sample processing can be accomplished in avariety of biochemical contexts and in a dramatically shorter timeperiod and in a manner that is easier for the operator. In fact, thepresent invention shortens tests that have previously taken 60 or 90 oreven 120 minutes to an intraoperative time frame such as 20 minutes orthe like. Embodiments of the invention overcome what may have beenpreviously considered a physical requirement, namely, that manyparticular biochemistries involved simply required a long time.Embodiments of the invention also permit coincidental processing of allsamples at once. Furthermore, by creating particular conditions withinthe system, the desired amount of chemical interactions can beaccomplished in a far shortened timeframe. In embodiments, the inventionacts to replenish a microenvironment on an exterior sample area of asample so that binding or more generally, other interaction, can occurmore rapidly. Embodiments of the present invention overcome the longerbinding times previously perhaps taken as a physical constant.Embodiments realize that by acting in a manner to replenish amicroenvironment, not just move fluid on a sample, can significantlyshorten the time needed for a particular amount of interaction. Ratherthan using a completely new application of reagents or the like, thepresent invention acts in a manner where the microenvironment isreplenished and a shortened interaction is achieved. Some embodiments ofthe invention achieve this by removing, perhaps mixing, and reapplyingthe same fluid so that the fluid and the substance in themicroenvironment immediately adjacent the sample is not depleted.Embodiments also provide systems for reading and storing consumableinformation such as many insist from reagents, wicking elements, and thelike.

4. BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a depiction of outer view of a self contained systemaccording to one embodiment of the invention.

FIG. 2 is a conceptual schematic depiction of one embodiment of a sampleprocessing system.

FIG. 3 is a depiction of an enlarged view of a bounded fluidicenvironment such as in between two slides.

FIGS. 4A-4D show a depiction of surface movement sequences such as inone embodiment that act to eliminate and replenish a fluidic substance.

FIG. 5 is a diagram of some representative antibody binding profiles.

FIG. 6 is a depiction of some rapid sample processing protocol steps andtimings.

FIG. 7 is a cut away depiction of one embodiment of a rapid sampleprocessing system.

FIG. 8 is an exploded view of the embodiment of the rapid sampleprocessing system in FIG. 7.

FIG. 9 is a depiction of a reagent magazine and cartridge showing usewith various substances.

FIG. 10 shows a depiction of an instrument view of a rapid sampleprocessing system showing certain structural elements in anotherembodiment.

FIG. 11 shows a depiction of a side view of a slide movement system inan open position for the embodiment in FIG. 10.

FIG. 12 shows a depiction of a side view of a slide movement system in adispensing position for the embodiment in FIG. 10.

FIG. 13 is a depiction of a side view of a slide movement system in apartially closed or partially open position for the embodiment in FIG.10.

FIG. 14 is a depiction of a side view of a slide movement system in aclosed position for the embodiment in FIG. 10.

FIG. 15 is a depiction of a side view of a slide movement system in atilted position for the embodiment in FIG. 10.

FIG. 16 is a depiction of a perspective view of a slide movement systemin an open position for the embodiment in FIG. 10.

FIG. 17 is a depiction of a perspective view of a slide movement systemin a closed position for the embodiment in FIG. 10.

FIG. 18 is a depiction of a close up view a slide movement system in aclosed position for the embodiment in FIG. 10.

FIG. 19 is a depiction of a perspective view of one embodiment of alinear reagent magazine.

FIG. 20 is a depiction of a perspective view of a different embodimentof a linear reagent magazine with attached primary antibody cartridge.

FIG. 21 is a depiction of an exploded view of an embodiment of a primaryantibody cartridge.

FIG. 22 is a depiction of an exploded view of an embodiment of a linearreagent magazine.

FIG. 23 is a depiction of the operation of an embodiment of a substancedispenser as may be included in either an antibody cartridge or areagent magazine.

FIG. 24 is a depiction of a cut away view of an embodiment of asubstance dispenser as may be included in either an antibody cartridgeor a reagent magazine.

FIG. 25 is a depiction of a perspective view of one embodiment of aperpendicular wicking cassette.

FIG. 26 is a depiction of a cutaway view of the embodiment of theperpendicular wicking cassette shown in FIG. 25.

FIG. 27 is a depiction of a perspective view of one embodiment of aparallel wicking cassette.

FIG. 28 is a depiction of a corner perspective view of the embodiment ofthe parallel wicking cassette shown in FIG. 27.

FIG. 29 is a depiction of a cutaway view of the embodiment of theparallel wicking cassette shown in FIG. 27.

FIG. 30 is a depiction of a corner cutaway view of the embodiment of theparallel wicking cassette shown in FIG. 27.

5. MODES FOR CARRYING OUT THE INVENTION

As mentioned earlier, the present invention includes a variety ofaspects, which may be combined in different ways. The followingdescriptions are provided to list elements and describe some of theembodiments of the present invention. These elements are listed withinitial embodiments, however it should be understood that they may becombined in any manner and in any number to create additionalembodiments. The variously described examples and embodiments should notbe construed to limit the present invention to only the explicitlydescribed systems, techniques, and applications. Further, thisdescription should be understood to support and encompass descriptionsand claims of all the various embodiments, systems, techniques, methods,devices, and applications with any number of the disclosed elements,with each element alone, and also with any and all various permutationsand combinations of all elements in this or any subsequent application.

The present invention can be understood by reference to the detailedfigures and description set forth herein. Embodiments of the inventionare discussed below with reference to the Figures. However, thoseskilled in the art will readily appreciate that the detailed descriptiongiven herein with respect to these figures is for explanatory purposesas the invention extends beyond these limited embodiments.

Referring to FIGS. 1, 2, and 10-19, it can be understood thatembodiments of the invention may present a self contained system (56)perhaps with a system enclosure (60) that can achieve a method of rapidsample processing. In general, the system may involve obtaining a sample(1), placing that sample in a sample processing system (2), and thenautomatically processing that sample (1) by operation of the system. Thesystem operator or other person can select an appropriate biochemicaltest sequence perhaps through a computer or perhaps touch screen display(57) or the like and the sample processing system (2) can be configuredas or can include an automatically sequenced test processor (3). Thesystem operator or other person can easily insert multiple reagentmagazines and perhaps single primary antibody cartridges for theselected sequences. In embodiments, the sample processing system (2) caninclude an automatically sequenced biochemical test processor (3), anautomatically sequenced histochemical test processor, an automaticallysequenced cytochemical test processor, or the like so that it may act toaccomplish a particular type of test not previously able to beaccomplished in or perhaps merely desired to be accomplished in a rapidmanner.

The sample processing system (2) can act to subject at least a portionof an exterior sample area (4) of the sample (1) to some appropriateinteraction. By permitting this interaction, the sample processingsystem (2) may be configured to cause or to permit the placement of asubstance (5) in the vicinity of the sample (1). Throughout the testsequence, substances are removed and held for disposal. As a result ofthe substance (5) placed on the sample (1), the sample processing system(2) can cause an appropriate interaction and thereby provide a detectionindication. This detection indication may by caused by the presence of aspecific type of biological substance within the sample (1). Asmentioned above the sample processing system (2) can be used in even anoperative environment. As such, it may be appropriate to make the sample(1) a thin biologic sample or the like. This sample may be placed on oneor more thin biologic sample holders (6).

It may also be desired to accomplish a coincidental or parallelprocessing of a variety of samples at once. As such, the sampleprocessing system (2) may have multiple sample holders. These sampleholders may facilitate establishing a sample (1) on a surface (7). Thissurface (7) may be a substantially planar surface so that the sample islaid flat for easier interaction. In instances where the sample (1) is athin biologic sample, the sample (1) may be placed on a slide such as amicroscopic slide (8). In this instance, the sample processing system(2) may include a microscopic slide sample holder (9). A microscopicslide sample holder (9) may facilitate the placement of may aid inapplying an appropriate substance (5) in the vicinity of at least aportion of an exterior sample area (4) of the sample (1). Byestablishing a sample (1) on a microscopic slide (8) traditionalstaining and analysis can be conducted, albeit in a shortened timeframe.

With respect to coincidental processing, it should be understood thatindividualized biological samples may be run with disparate, samplespecific biochemical test sequences. The automatically sequenced testprocessor (3) may act to automatically correlate a plurality ofdisparate, sample specific biochemical test sequences withindividualized biological samples. Through this action the sampleprocessing system (2) or perhaps the automatically sequenced testprocessor (3) may be considered as having a correlation element (67). Byknowing which sample is on a particular sample holder, the system canassure that the proper sequence is run with proper sample.

Through not only the proper software or firmware but also appropriatehardware, the system can act automatically substantially coincidentallyconduct at least a portion of biochemical test sequence. Bysubstantially coincidentally conducting actions, the system can causethose actions to occur and nearly the same time for more than onesample. This may of course encompass all of biochemical test sequence.It may also, encompass specific aspects such as: subjecting a sample toa substance, incubating a sample in this substance, eliminating orperhaps transiently eliminating the substance, reapplying thissubstance, withdrawing a substance, and even achieving or accomplishingthe ultimate desired results. Thus, by appropriate programming andperhaps use of appropriate mechanisms, the system can be considered ashaving a variety of substantially coincidental elements such as an:automatic substantially coincidental substance mixer, automaticsubstantially coincidental substance application element, automaticsubstantially coincidental paired sample substance application element,automatic substantially coincidental incubation element, automaticsubstantially coincidental substance transient elimination element,automatic substantially coincidental fluidic substance transientelimination element, automatic substantially coincidental capillarytransient elimination element, automatic substantially coincidentalsubstance reapplication element, automatic substantially coincidentalsubstance withdrawal element, substantially coincidental automaticallysequenced biochemical test processor, or the like. In instances wherewithdrawing a substance is accomplished by wicking, the system can beconsidered as having a substantially coincidentally wicking element andthe like. This may be a capillary action substance withdrawal elementwhen capillary action is used such as in a wicking situation. The systemmay be configured for fully coincidental actions as well. As can beunderstood from the figures, specifically considering FIG. 13 and themechanical design presented whereby through a common hinged movementelement (47), fully coincidental processing can be conducted for all thepairs of slides on that particular holder. Thus, by appropriateconfiguration, the plurality of samples can be configured forcoincidental processing. Furthermore, the automatically sequenced testprocessor (3) AP configured as a fully coincidental automaticallysequenced biochemical test processor.

As mentioned, at least a portion of an exterior sample area (4) of thesample (1) may be subjected to an appropriate substance (5). Thesubstance (5) may be any appropriate reactive or even a non-reactivesubstance. To the presence of an appropriate interactive or perhapsreactive substance may facilitate actions so that detection can occur.In many instances, a substance (5) may be a fluidic substance (10). Bysubjecting the sample (1) to at least one fluidic substance (10)interaction such as antibody binding, staining, or the like may occur.Naturally, the fluidic substance (10) may be an appropriate reactivesubstance or perhaps an appropriate fluidic reactive substance. Thus,the sample processing system (2) may include some type of substancesource, such as a fluidic substance source (11) as part of a preselectedbiochemical test sequence. The sample processing system (2) may alsoautomatically cause actions such as by the fluidic substance source (11)that place fluidic substance (10) on a sample (1). In embodiments of theinvention, this action may be conducted through the use of capillaryaction and thus a sample processing system (2) may capillarly subject asample (1) to an appropriate substance (5). In some populararrangements, the fluidic substance (10) may be a liquid substance. Thisliquid substance may of course be a solution, a suspension, or any othertype of substance. In other embodiments, the invention can even beadapted to a nonliquid fluidic substance such as a gaseous substance.

Once the substance has been placed on a sample (1), the automaticallysequenced test processor (3) may be programmed and may act to permit asample (1) to be incubated in the presence of a substance (5). Thisprogramming may act as an incubation element (12) within the sampleprocessing system (2). An automatically sequenced test processor (3) mayact to accomplish the step of incubating the sample (1) in the substance(5) for some period of time after it also accomplishes the step ofsubjecting the sample of (1) to the substance (5). In some embodiments,the sample processing system (2) can accomplish incubation an unelevatedtemperature, such as room temperature or the like. Naturally,embodiments may act to heat the sample or the like and the temperaturemay actually be increased by some amount. For temperature sensitivesubstances, embodiments of the sample processing system (2) may notsignificantly externally heat the sample or the substance (5) and thusthe sample processing system (2) may contain an unelevated temperatureincubation element (13) by causing, whether through programming or thelike, the sample (1) to incubate in the presence of a substance (5)without a significantly elevated temperature.

Of course, as the substance changes, the sample changes, or the processchanges, the automatically sequenced test processor (3) within thesample processing system (2) may be differently programmed to utilizediffering incubation periods for differing substances, differingsamples, or the like. Depending upon the substance or samples involved,it is also possible for the automatically sequenced test processor (3)to even utilize no incubation period. This may be appropriate ininstances where there is sufficient interaction in the time period wherethe substance (5) is placed and removed from a sample (1). It may alsobe appropriate for certain substances such as a buffer substance. Insuch instances, the buffer substance may be applied and relativelyimmediately removed with no significant incubation or delay period. Bythe term relatively immediately removed, it should be understood thatwhile there may be pauses or the like that may be incidental to themechanics or other processing aspects of the automatically sequencedtest processor (3), no substantial delay may occur and thus nosignificant incubation period may exist for particular arrangements.

Naturally, the amount of incubation can vary. Significant in someembodiments of the present invention is the possibility that incubationcan be greatly shortened as compared to prior techniques. Incubation mayalso be conducted in a sequence of partial incubation events. In somesuch partial incubation events it may be arranged such that theautomatically sequenced test processor (3) may act to partially incubatea substance for less than or equal to a variety of times. These timesmay range from 90 seconds to zero seconds. Partial incubation eventswhich may be such as 90, 60, 35, 30, 22, 20, 15, 10, 5, 3, and even zeroseconds maybe applied. In such events the sample (1) may also besubjected to the substance (5) without significant disturbance. In thisundisturbed timeframe, an appropriate interaction, reaction, or otherprocess can occur in a more traditional sense. In embodiments of thepresent invention, the amount of interaction can be far greater thanwould have normally occurred in the selected timeframe. Perhaps evenmore significantly, partial incubation can occur in time frames that arenow dramatically shorter than previously understood as possible for thedesired amount of interaction. Combining the partial incubationsequences, a total incubation time can thus be dramatically shortened.Again, through action of embodiments of the invention, the totalincubation of a particular chosen substance can even be for a total timeof less than or equal to about 300, 250, 200, 150, 20, 16, or even 10seconds as part of a selected biochemical, histochemical, cytochemical,or other appropriate sample process.

At the end of specific substance portions of the test sequence, theparticular substance involved is typically withdrawn from the sample.Especially when substances are reactive, this can present a hazardousmaterial handling issue. Embodiments of the present invention cancollect such spent substances in an enclosure for ultimate disposal. Foroverall simplicity and safety, snap in and snap out elements can be usedto aid the operator.

Embodiments of the invention can act to greatly shorten the chemistrytimes previously thought by some to be unchangeable constants. Throughappropriate programming, the automatically sequenced test processor (3)may act to initially permit an interaction between the sample (1) and anappropriate fluidic reactive substance. In situations as may beappropriate to many immunohistochemical tests, the system can beconfigured to permit a chemical interaction or even a chemical reactionto occur between the sample (1) and some substance such as an antibodysubstance (14) in a shortened time frame. These chemical interactions orperhaps reactions may take a variety of forms and may includeinteraction such as is present when an antibody binds to a particularcellular or other structure.

In embodiments of the invention, the system may be configured to act tofirmly confine and perhaps restrain the fluidic substance in thevicinity of a sample (1). This may create a bounded fluidic environment(15) or a restrictively confined fluidic environment (17). By firmlyconfining the environment, a fluid or other substance may be confined bysomething exhibiting a significantly greater resistance to movement thanfluid itself. This may be a rigid material or perhaps something that ispliable. In some arrangements a sample processing system (2) may beconfigured so that it establishes a bounded fluidic environment (15) inthe vicinity of the exterior sample area (4). This bounded fluidicenvironment (15) may be established through some type of fluidicboundary element (16). A fluidic boundary element (16) may actually bearranged and configured to permit the bounded fluidic environment (15)to exist in the vicinity of the sample (1). By acting to establish abounded fluidic environment (15) in the vicinity of the exterior samplearea (4), the system may serve to provide an environment within which anappropriate reactive substance (5) may be placed. Furthermore, thebounded fluidic environment (15) may serve a variety of purposes. First,it may act to limit the amount of fluidic substance (10) that is usedfrom a source such as the fluidic substance source (11). This may serveto conserve what may prove to be a very expensive substance. Inaddition, the bounded fluidic environment (15) may serve to facilitatean appropriate action on the fluidic substance (10).

In embodiments, the bounded fluidic environment (15) may be configuredto cause or permit a firmly or otherwise restrictively confined fluidicenvironment (17) in the vicinity of at least a portion of the sample(1). By presenting a restrictively confined fluidic environment (17),the sample processing system (2) may present a fluidic environment thatenhances processing. In some embodiments a sample processing system (2)may include a multidirectional fluidic confinement element (18) that canact in more than one direction. It should be understood that this maynot be merely a multidimensional confinement element, but rathermultidirectional in that the directions may even be within the samedimensional context such as when binding on a top and a bottom, perhapsconsidered a single dimension, the Z-axis.

In some embodiments, the multidirectional fluidic confinement element(18) may be rigid surfaces, and perhaps even a pair of rigid surfaces.It may also be configured as an opposing surface multidirectionalfluidic confinement element (19). This opposing surface multidirectionalfluidic confinement element (19) may have two surfaces that oppose eachother and thus confine a fluidic environment. In one embodiment,opposing microscopic slides (8) can be used to confine the fluidicenvironment. As shown, configurations may arrange samples in opposingpairs. These opposing pairs made me think of adjacent opposing pairs asshown in a lined arrangement in the figures. As shown in FIGS. 4A-4D,opposing microscopic slides (8) may act so that a small dimensioned areamay be present between the two microscopic slides (8). In this oneembodiment it can be understood how the restrictively confined fluidicenvironment (17) may serve to establish a multidirectional restrictivelyconfined fluidic environment in the vicinity of at least a portion of anexterior sample area (4). By establishing an opposing surfacemultidirectional restrictively confined fluidic environment in thevicinity of at least a portion of the exterior sample area (4), thatexterior sample area (4) may be preferentially subjected to anappropriate fluidic substance (10).

In some embodiments, the multidirectional fluidic confinement element(18) may be configured as an at least three directionally confinedfluidic confinement element (20). This may be achieved through the useof an opposing surface multidirectional fluidic confinement element (19)such as shown with two microscopic slides (8) and an additionalconfinement direction, perhaps at the end of a microscopic slide (8) orat its label element, hydrophobic element, hydrophobic label, or thelike. Naturally additional directional confinement can be provided.Confinement can also be accomplished through the use of appropriatematerials such as by using a hydrophobic material or the like. As butone example, it can be understood that by using a label that ishydrophobic, certain fluidic substances (10) may actually becomeconfined in yet another direction. Confinement can cause at least threedirections of confinement and thus can present an at least threedirectionally confined fluidic confinement element (20). Naturally itshould be understood that a multidirectional fluidic confinement element(18) may act to establish a restrictively confined liquid environment ormaybe even a restrictively confined gaseous environment.

As mentioned earlier, a variety of substances can be used in order toachieve the desired results. As such, a sample processing system (2) mayinclude a substance source (21) that acts as a particular type ofsubstance source. A great variety of substances can be used and thus thesubstance source can serve as a histochemical process substance source,a cytochemical process substance source, an organic substance source, acytologic substance source, and even on a biomolecular substance source.Even with more specificity, a particular substance that is appropriateto particular tests can be used and thus a substance source can alsohave different types of substances including but not limited toreagents, primary antibody substances, and secondary antibodysubstances, chromogens, cellular substances, counterstains,histochemical probes, cellular substance counterstains, first chromogencomponents, second chromogen component, monovalent antibody substances,multivalent antibody substances, histologic substances,immunofluorescence substances, immuno gold substances, immuno goldsilver enhanced substances, immuno cytochemical substances, immunohistochemical substances, fluorescent molecular substances, and evenbiologically specialized proteins. Further, substances produced by othersubstances can be used such as substances produced by antigenstimulation, substances produced by B-cell stimulation, B-cellstimulation produced proteins, immune response substances to otherelements, immune responses to antigens, immunoglobulins, and otherwise.Of course, a variety of stain substances can be used such as basophilicstains, acidophilic stains, hematoxylin stains, eosin stains,eosinophilic stains, H&E stains, Lee's stain substances, Mallory'sconnective tissue stain substances, periodic acid-Schiff stainsubstances, phosphotungstic acid hematoxylin stain substances, silverstain substances, Sudan stain substances, Wright's stain substances,Verhoeff stain substances, trichrome stain substances, geimsa stainsubstances, tristologic substances, cytologic substances, biomolecularsubstances, and even substances that contains any combinations of theabove. As should be understood, a substance source (21) may be part of asystem that includes an appropriate substance type of processor (22). Inthis fashion the system may include a histochemical processor, acytochemical processor, or the like. Furthermore the system can beconfigured to subject the sample to any of these substances.

An important aspect of embodiments of the present invention may be itsuse with particularly challenging substances such as low affinityantibody substances or perhaps low temperature antibody substances. Inthis manner, embodiments of the invention can serve to achieve resultswhere previously they may not have been practically possible. As oneexample, a low affinity antibody substance, such as any antibodysubstance that typically does not exhibit an acceptable percentage ofbinding within the previously understood time frames, can be used. Assuch, the sample processing system (2) may serve through its programmingor the like to subjects the sample (1) to a low affinity antibodysubstance. A type of low affinity antibody substance may even be asubstance that has not previously been effectively usable in automatedstaining devices. In addition to low affinity antibody substances, aheat sensitive antibody substance might also be used. While in somesystems such antibody substances may not have been used in the past, nowthey might be used to a greater degree. While some systems utilized heatto cause accelerated interaction within the previously acceptable timeframes, the present system may be able to be used with antibodysubstances that are heat sensitive. Thus some substances that may nothave been able to be used may now be usable. This may exist even if theaccelerated time frames are not available due to their intolerance toelevated temperatures and their low affinities. In some instances, anantibody that typically and traditionally bound less than about half ofits typical eventual amount in about 150, 180, or 240 seconds might beused. All this, of course, may even take place under normal temperatureconditions and thus the system may be used with an antibody substancethat traditionally takes longer than the mentioned time frames to bindabout one half of their typical eventual amount under normal temperatureconditions.

As mentioned earlier, a sample processing system (2) can act tofacilitate accomplishing a variety of different test sequences or testprocesses. Applying embodiments that permit rapid processing, the systemmay be configured to accomplish rapid immunohistochemistry,immunocytochemistry, in situ hybridization, fluorescent in situhybridization, chromosomal identification, staining, antigen retrieval,cytochemical, molecular chemical, epitope retrieval, or evenpretreatment processes. These different types of processes can also beapplied to a great variety of samples. Thus by obtaining a particulartype of sample and perhaps placing that sample in a sampleholder—perhaps such as a microscopic slide sample holder (9)—the sampleprocessor or perhaps the automatically sequenced test processor (3) canbe configured to process a variety of differing samples. These cansamples can be biologic, cellular, tissue, biopsy, carcinoma related,melanoma related, lymphoma related, margin testing related, epithelialcell, lymph node, undifferentiated tumor cell, pediatric cell, mohsmapping cells, h.pylori cells, an chronic villi tissue cells, neonatalherpes cells, proteomics cells, or other types of samples. As such, theprocessor can be any one of these types of processors throughappropriate programming to achieve a test and act on samples of the typementioned. The entire sample processing system (2) can provide adetection indication of the presence of some type of biologicalsubstance within the sample (1). This detection, may include a detectionindication of the presence of a carcinomic, tumor, phagocytic, lymphnode, transplant procedure, tumor differentiation, pediatric pathology,mohs mapping, margin, margin indicative, h.pylori diagnosis, therapeuticmarker, chronic villi tissue, neonatal herpes, virally, bacterially,infectious diagnostic, or just a molecular indicative type of substancewithin the sample.

A specifically important type of processing may be immunohistochemistryprocessing. As such, the sample processing system (2) may accomplish anyone of the particular types of processes involved inimmunohistochemistry. It may include an appropriate type ofimmunohistochemistry processor. When such a processor is appropriatelyconfigured, it may serve as an automatically sequenced test processor(3) and may be of any type mentioned. In general, biochemical processingmay include a great variety of types of chemical processing includingbut not limited to histochemical processing, cytochemical processing,immunohistochemical processing, and the like.

In some embodiments, a significant aspect may be the fact that thepresent invention can be configured to achieve rapid biochemicalprocessing. As such, the sample processing system (2) may include a fastbiochemical sample processor (24). This fast biochemical sampleprocessor (24) can act to accomplish the desired results and accomplisha completion time that is shorter than a traditional completion timeperiod. The traditional completion time can be considered as the time atwhich is presently understood as being the amount of time necessary inorder to achieve desired results for a given type of sample, given typeof process, given type of test apparatus, and/or a given type ofsubstance. For the same type of configurations, the present inventioncan achieve fast biochemical processing through the use of a fastbiochemical sample processor (24) which may be configured to achieve thesame desired results as might normally be chemically expected in longertime period. A fast biochemical sample processor (24) may act in lessthan a traditional completion time to achieve the same desired results.In some embodiments this may be achieved without elevated temperaturesand thus the system may act on an antibody substance to be in less thana traditional unelevated temperature binding time period.

As a fast biochemical sample processor (24) the system may be configuredto automatically achieve the biochemical test sequence in less than thetraditional completion time while still achieving the same desiredresults. This may even be an intentionally shortened reaction period inwhich the substance (5) may interact with the sample (1). In such aconfiguration a fast biochemical sample processor (24) may even act as abiochemically time shortened interaction element. This may serve as areduced detection time period process completion element. Embodimentsmay provide a detection indication in a reduced detection time period sothat it can be used in an intraoperative or other shortened timeenvironment. Naturally by presenting a method of rapid biochemicalprocessing it is possible that the present system may be used beyondenvironments that merely require shortened processing. It may also beused in instances where it is desirable to simply take less time. Ininstances where antibody substances are used to bind with a sample (1),a system may be configured to subject a sample to a reduced antibodybinding time period. This may be achieved by enhancing the interactionsubstantially along at least a portion of a bounded fluidic environment(15). It may of course also occur within a restrictively confinedfluidic environment (17).

By allowing enhanced interaction to occur for at least some period oftime, embodiments of the invention may permit rapid biochemicalprocessing. It should be understood that this rapid processing canmerely be something that is shorter than a recommended reaction period.The interaction need only occur in some shortened time frame as comparedto that traditionally accepted for a particular situation. Embodimentsof the sample processing system (2) may have reduced times as comparedto a traditional interaction, completion, reaction, or detection timeframe for a given situation. Perhaps surprisingly the present inventioncan achieve an acceptable level of interaction or reaction in less thanan anticipated to timeframe. Thus, although desired results weretypically not chemically expected until in longer time period, thepresent invention can present those same results in a shortenedtimeframe. This shortened timeframe may even be a less than arecommended reaction period and it may also be conducted as part of asimple automated biochemical test sequence.

By limiting a period within which an appropriate reactive substance maysubstantially be reacted, the present invention can serve to achieverapid results, namely, results in less than a traditional completiontime period. This timeframe may be an intentionally shortened reactionperiod. Embodiments of the invention made provide a detection indicationin the presence of the specific type of biological substance in areduced detection time period and may utilize a reduced reaction timeperiod element, a reduced interaction time period element, or perhaps areduced binding time period interaction element (25). This reduced timeperiod interaction element (25) may be programming such as containedwithin an automatically sequenced test processor (3) in the sampleprocessing system (2) or it may be present as hardware or firmware. Thereduced time process completion may be a reduced time process completionelement (26) and may be included in the sample processing system (2) aswell as a reduced detection time period process completion element. Byutilizing a reduced time period in whatever context, embodiments of thepresent invention can achieve the results even perhaps with unelevatedtemperature conditions. Thus, embodiments of the invention may utilize areduced unelevated temperature interaction time period for a particularsubstance.

In some embodiments, the amount of interaction may be an appropriateamount. In situations such as the binding of an antibody to a sample(1), embodiments of the invention may accomplish a significantpercentage of a traditionally accepted total amount of unelevatedtemperature antibody binding in a reduced time period. These significantpercentages may be percentages such as greater than or equal to about70, 80, 90, 95, 98, perhaps substantially all, or even 100% of atraditionally accepted total amount of unelevated temperature antibodybinding. A qualitative amount and time frame can also be provided suchas embodiments which provide a detection indication in less than orequal to about a visiting outpatient, an intraoperative procedure timelimit, or perhaps even the College of American PathologistsIntraoperative Guideline amount of times. By achieving results in thesemore general contexts, the present invention can offer systems that canbe used more effectively by doctors and more effectively for patients.The present invention may thus be appropriate for use in an operatingroom time constraint environment or the like. It may even permit use ina surgery time constraint environment or the like. Quantitatively,embodiments of the invention may provide a detection indication in lessthan about the aforementioned 60, 45, 30, 20, 15, 12, and even 10 minutetime frames. Aspects of the system such as the automatically sequencedtest processor (3) and such may be configured to serve as a reducedhistochemical detection time period process completion element. Thesemay be configured to provide a detection indication in less than any ofthe previously mentioned time frames. A system may also be configured touse interaction times that are less than about 75%, 50%, 30%, 23%, oreven 18% of a traditional unelevated temperature interaction time frame.This may also apply in situations with elevated temperatures as well. Asystem may also provide a completion element configured to provide theaforementioned detection indication time frames and made provide ahistochemically time shortened interaction element with theaforementioned time frames. As mentioned earlier, the system may providean indication in less than or equal to about 500, 400, 300, 240, 180,150, or perhaps even less than or equal to about 120 seconds times. Thismay occur for substances that cause about 50% or perhaps 80% of theirtraditionally accepted total amount of unelevated temperatureinteraction in longer than about 90 for the 50% amount or perhaps 660seconds for the 80% amount.

In order to achieve a shortened timeframe or interaction aspects, thesystem may include activities that cause a forced action within afluidic environment. The system may act to apply a motive force in thevicinity of the sample. This may occur through a motive force element(23). This motive force may be applied substantially along at least aportion of the restrictively confined fluidic environment (17). Byautomatically applying a motive force, the automatically sequenced testprocessor (3) may act at appropriate times. This application of a motiveforce may, in some embodiments, initiate a fluid wave. By affirmativelyinitiating a fluid wave, fluid within the restrictively confined fluidicenvironment (17) may be moved. The affirmative initiation of a fluidwave may also occur in a restrictively confined fluidic environment(17). All this may occur at time when the fluid needs or will beenhanced by being replenished in some fashion.

In some embodiments the automatically sequenced test processor (3) mayact to affirmatively initiate an oscillatory fluid wave, that is, afluid wave that may occur and move back-and-forth multiple times. Thesemay or may not be of a regular nature and may or may not have pauses inbetween. Through such a program, the system to include subroutines orthe like that may serve as an oscillatory fluid wave element (27).Naturally, it should be understood that the oscillatory fluid waveelement (27) need not be included. In some embodiments the system mayinclude a general fluid wave element. This fluid wave element may merelycause some sort of fluid wave to occur within a bounded fluidicenvironment (16) for at least some period of time period. A fluid waveelement may act within an area such as the restrictively confinedfluidic environment (17), and thus the system may cause fluid motion inthis environment. As explained later, in some embodiments this mightremove a fluid from the fluidic environment and then may even reapplythe fluid to that fluidic environment. The system may also act toautomatically substantially stop the fluid wave such as by removing themotive force that causes the wave in the first place.

As mentioned earlier, one of the aspects of an embodiment of theinvention may counter a depletion of a substance (5) as it interactswith the sample (1). As shown in FIG. 12, a sample can be subjected tothe substance (5) through some type of fluidic environment. This fluidicenvironment may be a restrictively confined fluidic environment (17).Within any type of fluidic environment, restricted or not, there may becontained a microenvironment (28) which may exist immediately next tothe sample (1). This microenvironment (28) may also be immediatelyadjacent or next to the sample (1). A microenvironment many containelements of the substance (5) which actually interact with the sample(1). When elements of substance (5) become depleted, the amount ofinteraction may slow down. An aspect of embodiments of the presentinvention may be the fact that this microenvironment (28) can bereplenished without replacing the entire fluid. Specifically, as can beunderstood from FIGS. 4A-4D, by eliminating the fluidic substance (1)from within the microenvironment (28), the fluidic substance source (11)can be replenished and subsequently replaced. Through appropriatearrangements, the sample processing system (2) can include a sampleinterface microenvironment affirmative depletion avoidance element (29).This sample interface microenvironment affirmative depletion avoidanceelement (29) may be a combination of programming and perhaps hardwarethat acts to achieve the appropriate activity. This action may be assimple as merely accomplishing substantial mixing within the sampleinterface microenvironment (29). Interestingly, while air knifes and thelike have been used, these appear to have not achieved the level ofmixing necessary in the microenvironment (28) in order to affordsubstantially reduced process times as embodiments of the presentinvention can now achieve. In fact, existing systems (which may even useair knife systems) still retain the old processing times of an hour orperhaps even 90 or 120 minutes whereas the present invention affordssignificantly shorter process times—times that are less than anintraoperative 20 minute guideline.

To minimize substance usage, and to efficiently achieve the goal ofrapid processing, embodiments of the invention can act tonon-replacingly substantially refresh the substance (1) in themicroenvironment (28) adjacent the sample (1). This can occur in thesample interface microenvironment, by acting to transiently eliminatethe substance (5) from within the vicinity of the sample (1) and thenacting to reapply that same substance. As mentioned, substantialrefreshing can occur. Even in other systems that move a fluidicsubstance (10), such does not appear to occur as evidence by the factthat even those systems still have the slow process times and do not actto rapidly process the sample as in embodiments of the presentinvention. As shown in FIGS. 4A-4D, this can occur by moving the perhapsfirm surfaces (7) that define the bounded fluidic environment (15) orperhaps a firmly restrictively confined fluidic environment. By the termfirm, it should be understood to encompass both rigid or even pliableboundary elements. It can also occur by taking advantage of the effectsof capillary action as shown. Through such activity, the sampleprocessing system (2) or perhaps the automatically sequenced testprocessor (3) through the inclusion of a subroutine or programming orthe like can be considered as having a sample interface microenvironmentsubstance refresher element (30). This can achieve refreshing thefluidic substance (10) without replacing it.

Chemically, by refreshing the substance a high level of the desiredactivity can continually occur. As shown in FIG. 5 with respect to arepresentative substance, in this can a particular antibody substance,this can be understood. In this graphic depiction of what a traditionalaccelerated/heated antibody substance binding profile may be (58) slowbinding activity may likely be due to depletion of the antibodysubstance in the microenvironment (28). Thus the typical binding of anantibody substance can take in excess of 14 minuets to achieve 95% ofits eventual amount of binding. By acting, perhaps through the fivewaves or the like as shown to repeatedly refresh the substance (5),embodiments of the system can achieve the refreshed antibody substancebinding profile (59) shown. Through this refreshing, the curve can berepeatedly on its steeper portion and thus a higher rate of binding orother interaction can be achieved.

To achieve this transiently, such as in where the substance (5) isavailable for reapplication at some later point in time, embodiments ofthe system can establish a collected fluidic substance (36) as shown.Elimination of the substance (5) can also be substantial in that most isremoved even though some perhaps slight amounts may remain on or in thevicinity of the sample (1) or the surface (7). It should also be notedthat eliminating the substance (5) from the sample (1) is not the sameaction as the substance itself becoming depleted such as by the bindingor other desired interaction occurring.

Furthermore, the system may act to momentarily hold the substance (5)eliminated from the sample (1) such as in the location indicated for thecollected fluidic substance (36). This can occur without disturbancesuch as might let the collected fluidic substance (36) remix as a resultof it being eliminated from the sample (1) perhaps from turbulence orthe like. This holding can simply be a pause in operation and thus thesystem may be considered as having a collected eliminated fluid pauseelement (31). This pause element may act at least some times afteraction of the substance transient elimination element. The holding ofthe eliminated substance can occur for a variety of times, such as timesless than or equal to about 4 seconds, 3 seconds, 1.5 seconds, 1 secondsor even 500 ms. Through these times, the collected eliminated fluidpause element (31) may be configured to be a collected eliminated fluidpause element configures to hold for any of the above times. Asindicated in FIG. 6, multiple pauses can be used and thus the system mayhave a multiple pause collected fluid pause element (33). This can occurrepeatedly as well. In such a configuration, a sample processing system(2) can act to repeatedly momentarily hold a substance (5) eliminatedfrom the sample (1).

As mentioned, movement of a fluid in the vicinity of the sample (1) maybe desired for some embodiments. In this more general context, a sampleprocessing system (2) according to one embodiment of the invention maybe configured to include a sample interface microenvironment mix element(34). This sample interface microenvironment mix element (34) mayactually be a substance mixer that acts after the substance source hasdispensed its substance (5). This may act to mix of the substance duringat least some part of a process. It may act to accomplish substantialmixing within the sample interface microenvironment (28). In suchconfigurations, the system may act to accomplish substantial mixingwithin a particular area immediately adjacent to the sample (1). Thisarea, termed the sample interface microenvironment or more generally amicroenvironment (28) may be a variable thickness. For instance, inembodiments a sample interface microenvironment maybe next to the sample(1) to a depth of about of perhaps at least 20 μm from the sample (1).In some embodiments mixing may occur within smaller ranges as well.While at least a 20 μm microenvironment may be mixed in someembodiments, in other embodiments much smaller distances such as perhaps10 μm, 1 μm, 500 nm, 200 nm, 100 nm, 50 nm and even within at leastabout 10 nm may be the area of interest. Mixing or replenishment in sucha sample interface microenvironment (28) may be appropriate for aparticular applications or configurations. Interestingly, as understoodfrom the simple fact that existing systems do not afford such rapidprocessing, this microenvironment mixing does not occur in existingsystems.

In mixing the substance (5) within the microenvironment (28), the sampleprocessing system (2) can act with two different types of processes. Aninitial mixing may occur and then may be followed by a different type ofmixing. In one embodiment, the system may act to initially morefrequently and on a shorter time frame repeat a step of transientlysubstantially eliminating a substance from the sample (1). After theseinitial, perhaps more rapid steps, a second lower frequency or timeperiod of repeating may occur. As one example, initially the sampleprocessing system (2) may act with very little pause time in between theacts of eliminating and reapplying the substance (5). Following thisinitial action, the sample process system (2) may act more slowly toonly occasionally eliminate material. Initial multiple steps may causesubstantially more mixing and may permit the substance (5) to bereplenished more quickly perhaps through number of initial repetitionsof the step of transiently substantially eliminating the substance froma sample.

Referring to FIGS. 11 through 14, it can be understood how embodiment ofthe invention can serve to eliminate fluidic substance (10) from withinthe vicinity of sample (1). By including generally a substance transientelimination element (32), the sample processing system (2) may act toeliminate a substance (5) momentarily from within the immediate vicinityof the sample (1). Such a configuration may serve to transientlysubstantially eliminate an appropriate fluidic reactive substance fromthe sample (1). This may occur by substantially eliminating a fluidenvironment from within the immediate vicinity of the sample (1). In theembodiment shown, it can be seen that the system can act to decrease abounded fluidic environment (15) and thus pullback the fluidic substance(10) from in the vicinity of the sample (1). This may occur throughcapillary action such as by the movement of a surface (7), perhaps asurface such as the microscopic slide (8). As shown, in one embodimentthe microscopic slides (8) may be angularly moved apart and thus may actto decrease the restrictively confined fluidic environment (17). In thismanner the movement mechanism and perhaps the programming may serve toact as a decreasing restrictive fluidic confinement element (35). Thisdecreasing restrictive fluidic confinement element (35) may act totransiently substantially eliminate the substance from a sample (1). Bytransiently, that is in a manner consistent with an eventualreapplication, and substantially, that is such that a microenvironmentis also largely removed so that it may be replenished, eliminating thesubstance (5) from the sample (1), the system can achieve goals for thisembodiment. It may momentarily remove the substance (5) from the sample(1) in a manner such that the substance (5) may be reapplied to thesample (1) at some point in time. Furthermore, by substantiallyeliminating a substance (5) in the sample (1), the system may act toremove most of the substance (5) from in the vicinity of a sample (1).Naturally, some smaller amounts of substance (5) may remain, however, asignificant portion of the substance (5) be removed and collected forappropriate mixing or reapplication. Thus, embodiments of the system mayact to substantially eliminate the bounded fluidic environment fromwithin the immediate vicinity of the exterior sample area (4) of thesample (1).

As can be seen in FIGS. 4A-4D, collected fluidic substance (36) may beavailable within some proximity of the exterior sample area (4). Thiscollected fluidic substance (36) may then be reapplied to the sample (1)perhaps through action of a substance reapplication element (37). Thissubstance reapplication element (37) may act upon at least a portion ofthe transiently eliminated substance, perhaps most or all of thatindicated as the collected fluidic substance (36). By reapplying atleast a portion of the transiently substantially eliminated appropriatefluidic reactive substance, the sample processing system (2) act toreplenish the microenvironment (28) caused by a fluidic substance (10)so that this microenvironment (28) is no longer depleted of theparticular substance (5) of interest.

Similarly, a reapplication of the collected fluidic substance (36) mayoccur by increasing a bounded fluidic environment (15), that is, byincreasing the restrictions on such an environment and thus making itsmaller. By increasing the restrictively confined fluidic environment(17) the system may be considered as having an increasing restrictivefluidic confinement element (38). The increasing restrictive fluidicconfinement element (38) can reapply at least a portion of a transientlysubstantially eliminated substance to a sample (1) such as the collectedfluidic substance (36).

In some embodiments, capillary action can be used to move the fluid asdesired. By decreasing a restrictively confined fluidic environment (17)in the presence of particular substances and surface materials, thefluidic substance (10) may be pulled back by capillary action. Inembodiments, this can even occur at a speed that is greater than acapillary movement speed for a particular configuration. If the surfaces(7) such as the microscopic slides (8) are angularly pulled apart at aspeed that causes the fluidic substance (10) to move more rapidly thanit would move normally through capillary action without movement, thesystem may be considered as presenting a greater than a capillarymovement speed fluid movement element (39). Furthermore, the sampleprocessing system (2) can be configured to provide a capillarydisplacement element (40) and perhaps one that serves as a capillaryfluid displacement element. In addition, the capillary displacementelement (40) can serve as a capillary action substance applicationelement (41) by moving a fluid back on to a sample (1). Capillary actionmay exist with a capillary capable liquid, that is a liquid that exhibitcapillary action for a particular configuration or the like. Such aliquid may also include a liquid exhibiting surface tension or a surfacetension liquid.

Other types of movement are also possible, for instance, it is possibleto have a hydraulically displaced fluid in the vicinity of at least aportion of an exterior sample area (4). A system may include a hydraulicdisplacement element or the like. In general, if using a capillaryaction, a system can be considered as having a capillary fluiddisplacement element for displacing a fluid in the vicinity of at leasta portion of the exterior sample area (4). It may also be considered asacting to capillarly eliminate a fluid environment when it pulls afluidic substance (10) away from the sample (1).

Accomplishing movement at a speed greater than a capillary movementspeed for a particular configuration, the system can be considered aspresenting a fast fluid movement. This, of course, can include fastfluid application as well as fast fluid elimination. This may exist inany situation in which fluid movement occurs faster than a normalcapillary movement for a particular configuration, and thus the systemcan be considered as presenting a fast fluid movement element. Fastfluid movement can occur across the sample at speeds which may rangefrom at least equal to about 0.05 m/s, 0.1 m/s, 0.125 m/s, 0.25 m/s, 0.5m/s, and perhaps as great as at least about 1 m/s. Thus a fluid movementelement (42) may be configured to accomplish any of the mentioned fluidmovement speeds or other speeds as appropriate. In embodiments, a fluidmovement element (42) can accomplish a fluid movement in an amount oftime rather than a particular speed. For configurations as shown wherethe sample (1) is situated on a microscopic slide (8), an amount of timecould be a time such as less than or equal to about one second, one halfsecond, 400 ms, 200 ms, 100 ms, or even 50 ms.

By referring to FIGS. 11 through 14, it can be understood that a varietyof mechanical arrangements can be used to achieve the mentioned rapidsample processing. In one type of mechanical embodiment, multiplesamples (1) can be configured in an aligned arrangement. FIG. 11 showsthat an upper glass microscope slide (43) can be connected to an upperslide holder (44). Similarly, a lower glass microscope slide (45) can beheld by a lower slide holder (46). Both the upper slide holder (44) andthe lower slide holder (46) can be connected through some type of hingedmovement element (47). The hinged movement element (47) may act topermit some sort of angular movement between slide holders and thus theupper glass microscope slide (43) and the lower glass microscope slide(45). In a general sense, the hinged movement element (47) may simplycause some angular movement component between a first and a secondsurface, such as the microscope slides (8) or other types of surfaces.This movement can occur through use of a motor, perhaps a stepper motorunder computer control such as lower slide holder motor (48) and upperslide holder motor (49). As can be understood from the drawings, theupper and lower lance microscope slides (43 and 45) he serve as amovable firm fluidic boundary element. When some type of motive forceelement acts on these movable firm fluidic boundary elements, fluidmotion can occur and this may be considered as causing fluid movement ofthe movable firm fluidic boundary element. The hinged movement element(47) may thus serve as a hinged fluid wave element. Furthermore, thepair of slides (43 and 45) may be a proximally paired sample holder.

A hinged movement element (47) may serve as a first and second surfacemovement element. While the surfaces shown in the figures are actuallymicroscopic slides, it should be understood that the surfaces need notbe planar. They may be substantially planar or flat; they can be curvedas well. The surfaces may also be rigid surfaces, a pair of rigidsurfaces, or a pair of substantially planar rigid surfaces. In FIGS. 14and 4C, when the hinged movement element (47) is in a closed position,the upper glass microscope slide (43) may be in close proximity to thelower glass microscope slide (45). Through some operation whether it bethrough software, hardware, or perhaps firmware, the sample processingsystem (2) may be considered to include a close proximity surfacedisplacement element. This close proximity surface displacement elementcan serve to permit movement or displacement of one surface relative toanother while also permitting positioning at some point where thesurfaces are in close proximity to one another. In one embodiment, theinvention can be configured to displace the first surface relative toand in close proximity to the second surface, The surfaces of course maybe, but need not be, microscopic slides (8).

As mentioned earlier, a motive force element (23) can cause angular orother movement between a first surface relative to its second surface.Angular movement can be seen by comparing the movements shown in FIGS.11 through 14 and FIGS. 4A-4D. It can be seen that the sample processingsystem (2) may be considered to include first and second surface angularmovement element. This element can act to displace a first surfacerelative to and in close proximity to a second surface. While differentaspects of these movements are shown in FIGS. 11 through 14 and 4A-4D,it should be understood that the ultimate sequencing achieved by anautomatically sequenced test processor (3) can include many variationsof these movements. As shown in FIG. 6 a specialized sequence ofmovements can be achieved to accomplish a particular application thatsubjects the sample to a substance, transiently eliminates thatsubstance, mixes the substance, reapplies the substance, and ultimatelywithdraws the substance from a sample (1). As indicated, various steptimings and sequences can be achieved. For example as shown in stepthree and four, an initial sequence of ways to mix a primary antibodycan be achieved followed by a wave sequence that may permit significantincubation time periods in general. It can also be noticed through FIG.6 that an entire detection sequence can be achieved in less than 15minutes—a significantly reduced time period as compared to most existingsystems.

As shown in FIGS. 4A-4D it can be seen how the angular movement elementcan actually achieve elimination and reapplication of a fluidicsubstance (10). Viewing the sequence shown in FIGS. 4A-4C in the orderA-B-C as but one example of a sequence, it can be seen how closing theslides together can cause the sample (1) to be subjected to a fluidicsubstance (10). FIG. 4A shows how the two surfaces, in this casemicroscopic slides (8) can be in an open position. In this arrangement,the fluidic substance (10) is eliminated from in the vicinity of, and iseliminated from, an exterior sample area (4) of the sample (1). Asshown, this may occur through capillary action whereby the fluidicsubstance (10) is pulled back from the sample (1) perhaps by the naturaltendency of the fluid in such an arrangement.

FIG. 4B shows the microscopic slides (8) in an intermediate position. Ascan be understood, the fluidic substance (10) may be pulled along thearea in between a microscopic slide (8) and may pass across the exteriorsample area (4). FIG. 4C shows how in one embodiment the microscopicslides (8) may be moved to a closed position and are in close proximityto each other. First, it can be understood that the fluidic substance(10) may now be fully covering all appropriate areas between themicroscopic slides (8). Furthermore, in FIG. 4C it can be seen that themicroscopic slides may not actually be perfectly parallel to each otherwhen they are in the closed position. The microscopic slides (8) mayhave attached to themselves some type of identifier. This identifier,shown in FIG. 4A as labels (50) can cause spacing through their ownthickness. Shim elements (81) can also be used. These shim elements (81)may be interleaved so that each does not impinge upon an adjacent oneand thus double spacing if not desired. Thus the holders can serve asinterleaved proximally paired sample holders and may be interleavedabout a shim element. Furthermore, the shim element (81) or even thelabels themselves may also be a hydrophobic element to aid in firmlyconfining the fluidic environment. In instances where the identifiers orperhaps labels (50) are relatively thick, it is possible that themicroscopic slides (8) do not become fully parallel and the spacing mayeven be narrower at the other end. This is shown in FIG. 4C as onepossibility. Minimizing spacing can serve not only to reduce fluid butalso to permit the use of existing label arrangements—even if notoptimum.

The reverse sequence should also be understood. Considering FIGS. 4A-4Cin the order C-B-A, it can be understood how the fluid movement element(42) can act to not only apply fluid to the sample (1) (as in the A-B-Ccase) but also to eliminate the fluid from a sample (1) (as in the C-B-Acase). Again, with FIG. 4C depicting two surfaces in close proximity toeach other, when the surfaces are moved apart in an angular fashion,they may ask to eliminate fluid from the exterior sample area (4) of thesample (1). As shown in FIG. 4B as the surfaces or perhaps themicroscopic slides (8) are moved apart, the fluidic substance (10) maystart moving back toward the hinged movement element (47). As the twosurfaces continue to increase their angular movement, a full eliminationof fluid can occur. Ultimately it may be sufficient that the fluidicsubstance (10) is moved beyond the exterior surface area (4) of thesample (1) and is ultimately collected in some other location. Thus, ascan be understood how to FIG. 4A, collected fluidic substance (36) mayexist as a result of the motive force element (23) acting upon thefluidic substance (10). Once eliminated from an exterior sample area(4), the collected fluidic substance (36) can become mixed to somedegree and can be ready for reapplication.

Naturally a variety of spacings are also possible. In the embodimentshown in FIG. 4C, in the close proximity position, the surfaces may besubstantially parallel even though not perfectly parallel. The distancesbetween the two surfaces can vary based on the particular needs of thesubstance (5) or perhaps the sample (1). In instances in which thesample processing system (2) is configured for immunohistochemicalprocessing, it may be appropriate to use close proximity surfaces thatare separated by 100, 200, 250, and perhaps as little as at most about300 μm. Furthermore the separation (perhaps of at least one end) can beestablished by the identifier or perhaps label (50) thicknesses. Asshown in FIG. 4C, one end of the surfaces may be approximately two labelthicknesses apart. Similarly in situations where one of the microscopicslides (8) either does not contain a sample or has no label on it, themicroscopic slide (8) may be separated on one end by only one label oridentifier thickness.

Referring to FIGS. 8 and 16-18, it can be seen that a plurality ofsamples (1) may be held by a plurality of sample holders. In such anarrangement, it can be seen that the microscopic slides (8) may be heldin place by some sort of slide retention element, perhaps such as aslide retention spring (51). Of course, other arrangement are possible.In such an arrangement, multiple samples can be processedcoincidentally. For convenience, these multiple samples may be placedadjacent to each other and moved as one. In this configuration, thesample processing system (2) may have a sample holder that serves as amultiple, close proximity, substantially parallel or perhaps planarholder for a particular type of sample, a microscopic slide, or perhapsmerely surfaces.

The sample holder may also serve as a multiple, close proximity,substantially parallelly-oriented sample surface pair or perhaps even aproximally paired sample or surface as shown in FIGS. 10-18. Asmentioned with reference to FIGS. 4A-4D, it is possible to vary thespacing in order to alter the amount of fluid involved. As shown inFIGS. 3 and 4A-4D, this microenvironment (28) may be within a volumeimmediately adjacent a sample (1). By capillary or other action causingthe elimination or at least substantial elimination of the fluidicsubstance (10) (some fluidic substance may remain on a sample) thesystem can act to cause an adequate or even total removal of themicroenvironment (28). By eliminating the fluidic substance (10) andpulling it back into a collected fluid substance fluidic substance (36)the fluidic substance (10) may be refreshed and mixed.

A variety of fluid volumes may be used of course. In an embodiment andconfiguration where the system is designed for use to accomplishimmunohistochemistry with a microscopic slide, it may be appropriate tomove less than or equal to about 300, 225, or even 200 μl of a fluid.This movement may occur in at least a portion of the restrictivelyconfined fluidic environment and they occur by eliminating themicroenvironment (28). Thus, the sample processing system (2) may beconsidered as having elements configured to move less than or equal toabout any of the amounts of fluid mentioned earlier. Throughconsideration of particular substances and a range of samples, it may beappropriate to move a minimal amount of a substance (5) or perhapsfluidic substance (10). This minimal amount may be an amount thatpermits adequate replenishment of a microenvironment (28), or thatpermits adequate interaction between this sample (1) and the substance(5) in a selected process time period. In situations where there aredifferent or lesser demands on the time for processing it may bepossible to even further reduce a volume of fluidic substance (10) andstill permit adequate interaction between the sample (1) and thesubstance (5) in a desired time frame. As such the system may beconfigured to use a lesser amount of substance, rather than to minimizetime.

FIG. 6 is a table that shows that a variety of repetitious actions thatcan be accomplished in one example of a sequence. As shown, theautomatically sequenced test processor (3) can repeatedly mix orotherwise act upon a substance (5) or perhaps a sample (1). Referring tostep three, it can be noticed that a sequence may be used to initiallymix a primary antibody. Such mixing may involve, as but one example,three sequences where the fluidic substance (10) he pooled as acollected fluidic substance (36) and eliminated from the exterior samplearea (4). This may be held for a relative short time period (or no timeperiod at all) such as approximately 1.5 seconds. Similarly, during theinitial mix action the fluidic substance (10) may be held exposed to andreapplied to the sample (1) for some time (or again, or no time periodat all) such as in one example, two seconds.

Referring to step 4 in the exemplary sequence shown in FIG. 6, thevalues indicate that immediately following this initial fast wave actionthere may be a slower wave action with exposed incubation lastingperhaps as long as 22 seconds. This may be repeated, perhaps as shownfor six times or the like. While this represents but one particular testsequence, in general it should be understood that repeated actions canoccur. By its programming the sample processing system (2) or perhapsalso the automatically sequenced test processor (3) may be considered asincluding not only a repetitious action element but also perhaps in thisinstance a repetitious action fluid wave element. Through steps such asthe steps three and four as shown in FIG. 6, the system can act torepeatedly transiently substantially eliminating the substance from thesample (1). Naturally other repetitious action elements are possible,and these may include repetitious action elements such as: a repetitiousaction fluid wave element, as mentioned above, a repetitious actionsubstance elimination element, a repetitious action substancereapplication element, a repetitious action incubation element, or thelike. In instances where at least two repetitious actions arecontemplated, the system may be considered as having an at least twoaction repetitious action element, etc. The repetitious actions mayoccur regularly or perhaps irregularly. In the example shown in FIG. 6,this particular sequence of steps three and four involves two differentregular time intervals for at least a portion of the overall process.Thus, step three involves three repeats with a regular time interval andstep four involves six repeats with a regular time interval. In someembodiment the automatically sequenced test processor (3) may repeatedlyreapply at least a portion of a transiently substantially eliminatedsubstance.

The repeating activity may occur more than one, two, three, four orperhaps even five times. As shown in FIG. 6, it may occur three timesfor an initial mixing, six times for a primary antibody incubation andperhaps four times for an initial chromogen or perhaps two-stepsubstance mixing. Similarly, for a counterstain it may occur four timeswith an incubation or dwell time of perhaps four seconds as shown. Insome embodiments particularly those configured for immunohistochemistry,3, 6 and 9 repetitions may be appropriate for first antibody substances,chromogens, or counterstain substances. Through this action, it can beunderstood how the system can act to repeatedly and non-replacinglysubstantially refresh the substance in a microenvironment (28). Thismicroenvironment may be situated adjacent to the sample to permitshorter partial interaction time as well as a shorter overall processingtime in order to permit more rapid processing. Through this action thesystem can be considered as repeatedly causing a fluid wave within abounded fluidic environment (15). This can occur with a variety offrequencies including but not limited to the range of times present inthe ones specific test sequence shown in FIG. 6 as well as otherfrequencies. As may be understood from the variations shown for thespecific substance and sequences indicated in FIG. 6, the mixing canoccur with differing occurrences for differing substances. This is alsotrue of the elimination and reapplication steps. Furthermore the amountof time a collected fluidic substance (36) is held eliminated from theexterior sample area (4) vary as well.

As mentioned, the system may act to automatically withdrawal a substancesuch as a fluidic substance from within proximity to sample. This canoccur by using an absorbent material (72). This absorbent material maybe a wicking material. A substance withdrawal element (53) may beconfigured to withdraw substance from one or more samples. It may act tocome in transient contact at a location where fluidic substance exists.This transient contact location may be selected as a particularlydesirable location from which to withdraw a fluidic substance. Asmentioned the withdrawal of the spent substance may be through a wickingelement (73). The wicking element (73) can act to pick away a substancefrom it proximity of the sample. This wicking can exist by or because ofcapillary action and thus the wicking element may present a capillaryaction substance withdrawal element. Withdrawal of this substance from asample can be enhanced in some fashions. In embodiments, the system maybe configured to provide an enhanced withdrawal orientation element.This element may be an enhanced withdrawal orientation sample tiltelement (74). By establishing a substance withdrawal enhancementcondition for the sample (1), the system may facilitate withdrawal whilethe substance withdrawal enhancement condition is established. Thissubstance withdrawal enhancement element (74) can be both programmingand mechanical operation. In one embodiment, the system may act to tiltsample to an enhanced withdrawal orientation. This may involve orientinga surface to facilitate wicking of a substance, establishing a tiltedsurface, establishing an untilted surface, and establishing surfaces atat least about 22.5° , 30° , 45° , and perhaps 67.5° . Furthermore whensurfaces are angled with respect to each other a bisected angle betweenthe two surfaces, may be tilted. Again, this can occur at various anglesperhaps at least about 22.5° , 45° , and 90° .

FIG. 15 depicts a perpendicular absorbent wicking roll (52); FIGS. 7, 8,and to some extent 27 through 30 depict systems configured with aparallel substance withdrawal element (53) and having upper and lowerslide cameras (63 and 64). When any particular fluidic substance (10)has accomplished its function and is no longer needed, that substance(5) may be withdrawn. This withdrawal can occur in a variety of fashionsperhaps such as by automatically moving an absorbent material to aposition in the vicinity of the sample. Thus, system may have absorbentmaterial movement element (75). In the embodiments of the system, theabsorbent material movement element (75) may be a linear absorbentmaterial movement element that may permit an absorbent material to movealong a straight path forward and backward when needed. When extended,the absorbent material may make contact with at least some of thefluidic substance. In embodiments where the absorbent material iscontained within it confinement enclosure, the system may act toautomatically move the confinement enclosure and achieve the withdrawalof substance. When extended, it absorbent material may even be pressedpast a point of initial contact to assure adequate wicking. Thus, thesystem may contain an absorbent material substance pressure element (76)such as may be contained in programming of or hardware constituting amovement mechanism.

As mentioned earlier, the absorbent material may be encased inconfinement enclosure (75). This confinement closure (75) may beconfigured to substantially encase the absorbent material. Furthermore,in instances where multiple samples and perhaps varying amounts ofmultiple samples the system may be configured to establish a coordinatedexposed area of absorbent material that is appropriate for ananticipated amount of a substance to be withdrawn. By coordinating anabsorbent material parameter, be it width, length, type of withmaterial, thickness, or the like, the system can be assured of notreaching a saturation level—especially when adjacent samples areinvolved. Thus, the absorbent material may have a multiple samplesaturation coordinated parameter. The absorbent material exposed areamay be coordinated for an anticipated amount of substance.

As mentioned, the substance withdrawal element may be enclosed and maypresent a cassette. This cassette or other enclosure may be removablyengaged by some removable engagement element, perhaps a snaparrangement. The operation may be accomplished by merely extending andthen retracting the wicking cassette or other substance withdrawalelement. The substance be removed from any further interaction, and itmay be appropriate to withdraw the substance by some type of wickelement. This wick element may also act to capillarly withdraw thesubstance from proximity to the sample (1) upon the completion of atleast a portion of the process. As shown in FIG. 6, in onerepresentative test sequence, the withdrawal of the fluidic substance(10) can occur multiple times throughout the overall process. Forinstance, steps 5, 8, 11, 15, 18, 21, etc., indicate that at multipletimes a particular substance is withdrawn from the sample (1) as part ofthe overall test sequence. Withdrawal can also occur at times when nosubstance is supposed to be on a sample such as that shown in step 1. Atthis point in time this could be considered unnecessary however it mayserve to assure that the sample(s) are dry and ready to beginprocessing. As shown in FIGS. 7, 8, and 27 through 30, this cassette maybe arranged with a parallel major axis that is oriented with a pluralityof samples and thus the system may have a parallel major axisorientation element (76). As shown in FIGS. 15, 25, and 26, it may alsobe oriented in a perpendicular fashion, perhaps with a perpendicularmajor axis orientation element (77). While FIGS. 15, 25, and 26 show aperpendicular orientation element, FIGS. 7-8 and 27 through 30 indicatea parallel embodiment in which one cassette can be used. As shown, theupper and lower slide holders (49 and 48) may be arranged so thatmicroscopic slides (8) are configured in a row of pairs along a pivotaxis of a hinged movement element (47). This holder is one type oflinearly arranged plural sample holder. As such, when fluidic substance(10) is transiently eliminated it becomes a collected fluidic substance(36) closest to the pivot axis of the hinged movement element (47).Furthermore to facilitate withdrawal of the substance the upper andlower slide holders (49 and 48) may be tiled. At this point, a parallelwicking cassette can be moved into place to coincidentally withdraw thespent substance from within the vicinity of the samples (1). As shownthe substance withdrawal element (53) can be established so that theabsorbent material is configured in a median angular orientationrelative to the samples. In this arrangement, the major axis of theabsorbent material, that is the axis of the long dimension of itsexposed material is parallel to a plurality of linearly arrangedsamples. Furthermore, the cassette itself may be tilted to correspond toany tilt of the samples, perhaps by being aligned with a bisected angle.Thus the mechanical configuration (in this embodiment) may serve as amedian angular orientation element which orients the absorbent materialappropriately for withdrawal of the collected fluidic substance (36).

Regardless of whether the absorbent material (72) is used in a cassetteor other arrangement, embodiments may include multiple amounts ofabsorbent material (72) that may be sequenced so that in the series ofactions in any test or in separate tests, multiple events of withdrawingdiffering spent substances can occur. Thus, additional amounts of unusedportions of a larger amount of absorbent material (72) may be presentedto a location such as that of a collected fluidic substance (36) ormerely in the vicinity of the sample(1) or the samples (1). Toaccomplish this, embodiments can include an absorbent material sequenceelement (78). Through action of the automatically sequenced testprocessor (3), or more generally, the sample processing system (2), thesystem may act to automatically sequence the absorbent material so thatan unused portion is sequentially presented at appropriate times. Thismay occur through an absorbent material advance element that establishesan unused portion of absorbent material in an exposed position as partof the biochemical or other test sequence. In addition, embodiments mayadvance or sequence the material in appropriate amounts perhapsincrementally and so there may be included, perhaps through programmingor the like, an absorbent material multiple sample appropriateincremental advance element that acts to only sequence an appropriateamount based on how many samples were processed.

As shown in FIGS. 27 through 30, in one embodiment, a more commoncassette arrangement such as that with a rotation element, perhapsmultiple roller elements, can be used. Whether in an absorbent materialroller system or otherwise, embodiments may act to accumulate a usedportion separate from an unused portion. This may occur through a usedabsorbent material accumulator that may even rotate an element and rollsome of the absorbent material. Two rollers may be used so that one actsto unroll an unused portion of absorbent material perhaps through anunused portion roller (79), while another acts to roll-up a used portionof absorbent material, perhaps through a used portion roller (80). Thesemay be coordinated so that the unused portion roller acts when the usedportion roller does. A two reel absorbent material roller cassette canbe used perhaps so that the system can be considered as continuouslyrolling two reels of absorbent material, that is sequencing throughmaterial at appropriate times and perhaps even in appropriate incrementsuntil the entire cassette has been used.

As mentioned, the absorbent material can be advanced in multiple sampleappropriate increments. In embodiments such as shown in FIGS. 10 and 16through 18 that have upper and lower slide holders (49 and 48) arrangedin a row as one type of linearly arranged plural sample holder, thesample processing system (2) may be operated in a manner in which notall positions are used in all test runs. In such a manner, or otherwise,it may be appropriate to use an altered amount of absorbent material(72). For example, while a row of eight sample pairs is presented in theembodiment shown in FIGS. 10 and 16 through 18, perhaps only fivesamples may need to be run with three different tests. In such aninstance, three locations (two pairs and one run by itself) may beactioned. If perhaps ten withdrawal events are needed for each of thesetests (e.g., primary antibody, rinse, secondary antibody, rinse,chromogen 1, rinse, chromogen 2, rinse, counterstain, rinse) only 30singular withdrawal spots may be needed. Rather than using withdrawalprogramming for when a full sample holder is used, where 80 singularwithdrawal spots may have been needed, the system may automatically actto advance in multiple sample appropriate increments and thus conserveabsorbent material (72). Furthermore, since a cassette or the like mayhave only a finite amount of total absorbent material, the system cantrack the multiple sample appropriate increments and perhaps totalizethem through programming or the like that may act as a multiple sampleappropriate increment tracker element. It may even inform an operatorwhen it may be appropriate to replace an element such as a cassette orthe like. Referring to the above example, it may also be understood thata single sample can be run as well. In fact, through proper programmingand in conjunction with the mechanical system presented, theautomatically sequenced test processor (3) may present a single sampleprotocol automatically sequenced biochemical test processor.

In presenting an exposed, likely unused portion of the absorbentmaterial (72), embodiments may act to tension an unused portion of theabsorbent material in an exposed position. Thus can occur through anabsorbent material tension element perhaps such as through exposedmaterial rollers (81) or the like between which said unused portion istensioned as it is exposed to the spent fluidic substance. Two materialrollers may be used as shown, and these may also help to reduce anywicking into the unused material roll during nonuse, so that the nextrun still present fresh, fully absorbent material.

FIGS. 11 through 14 and 16, show that chemical interaction may occurwhere the surfaces are, or at least one surface is, in an untiltedorientation. By establishing an untilted surface, it may be easier tofacilitate reagent dispensing on a surface such as a microscopic slide(8) such as by extending and retracting a reagent container (55)relative to a desired position or the like. The substance (5) may thenbe dispensed perhaps drop-wise or in an appropriate metered amount. Attimes when it is desirable to withdraw a particular fluidic substance(10) from an exterior sample area (4) from proximity to the sample (1),it may be desirable to facilitate this withdrawal of the substance insome manner. This can be facilitated in a variety of ways. In oneembodiment this may include orienting a surface to facilitate thewithdrawal of the substance away from proximity to the sample. As shownin FIG. 15, this orientation may involve establishing a tilted surfaceor perhaps tilting the surface to facilitate the wicking away of thesubstance. This orientation, or perhaps tilting, can occur at a varietyof angles and can establish either an upper or lower surface at a givenangle or perhaps a bisected angle at a given angle relative tohorizontal. This bisected angle may actually be a line in between thetwo surfaces and may be oriented at a particular angle. As shown in FIG.15 in one embodiment the orientation may be the tilting of a bisectedangle (54) at an angle such as 45°. In this configuration, the surface,perhaps a lower glass microscope slide (45), may be established atperhaps greater than 30° or even 22½°. Similarly the upper surface suchas the upper glass microscope slide (43) may be established at adifferent angle perhaps 60° or 67.5°. In this fashion it can beunderstood that the withdrawal of the fluidic substance by somethingsuch as the perpendicular wicking roll (52) as shown in FIGS. 15, 25,and 26 or perhaps the more general substance withdrawal element (53)such as perhaps a wicking cassette or the like may be moved intoposition to withdraw a liquid that has collected closer to the hingedmovement element (47). Of course, 90° could be used for any surface orperhaps even for the bisected angle (54). In some configurations it maybe appropriate to establish a tilted bisected angle (54) between the twosurfaces while accomplishing the step of wicking or more generallywithdrawing a substance (5) from proximity to the sample (1). This mayalso occur with one surface untilted such as might be desired toeliminate some movement element or motor or the like.

An aspect of some embodiments may be the fact that the sample processsystem (2) may act substantially coincidentally on all samplescontained. From FIGS. 10, 16, 17, 18, and 19 it may be understood thatone appropriate configuration of the system may present a substantiallycoincident sample treatment element to which multiple samples areresponsive. In the configuration shown this may involve the multiplesample holders shown. Such a system may also involve the use ofindividual or location specific reagent containers (55). These maydispense from a side actuator button (61) as shown in FIG. 19, from atop actuator (62) as shown in FIGS. 20-24, or otherwise. As shown inFIG. 12, the reagent containers (55) can move or otherwise place reagenton each microscopic slide pair. Further, as may be understood from FIGS.20-24, the reagent containers (55) may be configured to include one ormore of a cartridge (65) or a magazine (66). The cartridge (65) may be acontainer for a single antibody substance or the like, perhaps such asthe primary antibody substance. The magazine may be a single containerfor multiple substances with multiple chambers and thus may present asingle container multiple chamber multiple fluidic substance magazine(69). It may also be configured in somewhat of a line and thus present alinear reagent magazine such as is shown in one embodiment. The systemmay include a single container multiple chamber multiple fluidicsubstance magazine whereby a single container may have multiple chamberswith in it so that multiple fluidic substances can be placed in thechambers. In one arrangement, a dispensement force element (68) may beused to release substance from the single container multiple chambermultiple fluidic substance magazine (69). By having at least twosubstance chambers, the system can act automatically to determine whichsubstance or interactive fluidic substance is appropriate and can thendispense that substance. All that may be necessary for the operator isthe place the single container multiple chamber multiple fluidicsubstance magazine (69) in the system. In order to facilitate processingthat differs by location, the system may be configured to includelocation specific sources. These location specific sources may even havea correspondence with sample of locations. By including multiplesubstance source magazines, as one type of a location specific multiplesubstance source, differing substances can be dispensed on one samplethroughout the course of an automated test sequence. In addition, thesystem may include location specific single substance sources as well.Thus the system may include a single container multiple chamber multiplefluidic substance magazine (69), a linearly disposed multiple substancesource (70), or even a primary antibody cartridge (71) as shown in FIGS.20 through 21. Thus, the system can act to automatically process sampleseven when differing substances are required.

In order to minimize operator requirements when multiple runs may occur,the multiple substance source may include functionally relatedsubstances. furthermore substances can have volumes relative to theirpredicted usages. in this manner consistent may include a functionallyrelated multiple substance source magazine, and even a relativesubstance use sized source. To facilitate dispensement of the substanceonto the sample, the multiple source may be arranged in a row and movedas appropriate to permit the selected substance to be dispensed. asshown in FIGS. 20 through 24, the multiple substance source a beinglinearly disposed multiple substance source (70). Since primary antibodysubstances are of particular value, a separate single substance source,perhaps a primary antibody cartridge (71) can be used. This can be aseparate item or, and shown in FIGS. 20 through 24, it may even snapinto the multiple substance source magazine.

Appropriate mechanisms and software can achieve sampling and otherprocessing on all samples coincidentally. Thus the samples may betreated virtually identically in terms of the process sequences whileyet having their own particular substance selections as may beappropriate. This may involve not only coincidentally acting all in allsamples but also substantially evenly subjecting all samples to anappropriate fluidic reactive substance such as at the same time or thelike. All actions may even occur coincidentally in some embodiments. Thesystem though its programming may be considered as including asubstantially coincident sample treatment element. This substantiallycoincident sample treatment element may also act substantially evenlywith respect to its movement actions.

As may be understood with reference to FIG. 9, it can be seen thatdifferent primary antibody substances and even different to othersubstances may be included for individual slide locations. Thus,different substances may be dispensed while yet permitting all samplesto be treated substantially evenly and perhaps coincidentally. Asmentioned earlier, reagents may snap into specific locations. Anoperator a snap in at least one source, or they may even snap in orsomehow otherwise detachably connect even a linear multiple reagentmagazine. Thus, the system can be considered as having a snap securablelocation specific substance source, a snap securable primary antibodysubstance cartridge, or even a snap securable linear multiple reagentmagazine.

As may be understood from the detachable reagent cartridges, magazines,and the wicking cassette as but some examples, the system may involveuse of at least one consumable biochemical process element. These may beconsumably responsive to the automatically sequenced test processor (3).In order to understand substance, material, or, more generallyconsumable availability, it may be advantageous to know both how much ofan item has been used and how much of an item is available. For this andother purposes, embodiments may include a consumable process alterableinformation memory element perhaps in this embodiment a consumablebiochemical process alterable information memory element (82). Thisconsumable biochemical process alterable information memory element (82)may be physically or otherwise associated with one or more consumablebiochemical process elements. Upon installing, snapping in or attachingthe consumable, a consumable biochemical process alterable informationmemory element detachable electrical connection may be established suchas by a plug, an electrical, or any other type of connection. Throughsome type of consumable biochemical process alterable information memoryelement detachable electrical connection direct communication to andfrom can be established. The automatically sequenced test processor (3)or other aspect of the sample processing system (2) may act throughsoftware or the like to query the memory and perhaps even to change itupon consumable usage or the like. As such the system may include aconsumable information query element (83) and perhaps even a consumableinformation change element (84). These are but examples of what may actas a consumable biochemical process element. In one embodiment, thesystem may include an electrically erasable and programmable memoryelement attached to a consumable biochemical process element. Thisconsumable biochemical process element may be any items that may beconsumed or worn out in use of the system. For instance, using thesubstance withdrawal consumable biochemical process element such as thewicking cassette as one example, this item may have an electricallyerasable and programmable memory element attached to it. In thisinstance this may be considered a wicking element electrically erasableand programmable memory element (85). Similarly, for a fluidic substancecontainer such as the cartridge (65) or the magazine (66), there may bea fluidic substance container electrically erasable and programmablememory element (86) attached to either or both of these. Thusconsumables such as these can be more easily handled and monitored.Finally, whether with or without the memory and information elementsjust mentioned, the system can be configured with a network connectionor other communication modality to interface with a laboratoryinformation system or the like. This can be considered a laboratoryinformation system interface element (87).

As can be easily understood from the foregoing, the basic concepts ofthe present invention may be embodied in a variety of ways. It involvesboth processing techniques as well as devices to accomplish theappropriate processing. In this application, the processing techniquesare disclosed as part of the results shown to be achieved by the variousdevices described, as described steps, and as steps that are inherent toutilization as are simply the natural result of utilizing the devices asintended and described. In addition, while some devices are disclosed,it should be understood that these not only accomplish certain methodsbut also can be varied in a number of ways. Importantly, as to all ofthe foregoing, all of these facets should be understood to beencompassed by this disclosure.

The discussion included is intended to serve as a basic description. Thereader should be aware that the specific discussion may not explicitlydescribe all embodiments possible; many alternatives are implicit. Italso may not fully explain the generic nature of the invention and maynot explicitly show how each feature or element can actually berepresentative of a broader function or of a great variety ofalternative or equivalent elements. Again, these are implicitly includedin this disclosure. Where the invention is described in device-orientedterminology, each element of the device implicitly performs a function.Apparatus claims and method or process claims are support regardless ofthe nature of the discussion.

A variety of changes may be made without departing from the essence ofthe invention. Such changes are also implicitly included in thedescription. They still fall within the scope of this invention. A broaddisclosure encompassing both the explicit embodiments shown, the greatvariety of implicit alternative embodiments, and the broad methods orprocesses and the like are encompassed by this disclosure. It alsoshould be understood that language changes and broader or more detailedclaiming may be accomplished at a later date. With this understanding,the reader should be aware that this disclosure is to be understood tosupport as broad or narrow a base of claims as deemed within theapplicant's right and may be designed to yield a patent coveringnumerous aspects of the invention both independently and as an overallsystem.

Further, each of the various elements of the invention and claims mayalso be achieved in a variety of manners. Additionally, when used orimplied, an element is to be understood as encompassing individual aswell as plural structures that may or may not be physically connected.This disclosure should be understood to encompass each such variation,be it a variation of an embodiment of any apparatus embodiment, a methodor process embodiment, or even merely a variation of any element ofthese. Particularly, it should be understood that as the disclosurerelates to elements of the invention, the words for each element may beexpressed by equivalent apparatus terms or method terms—even if only thefunction or result is the same. Such equivalent, broader, or even moregeneric terms should be considered to be encompassed in the descriptionof each element or action. Such terms can be substituted where desiredto make explicit the implicitly broad coverage to which this inventionis entitled. As but one example, it should be understood that allactions may be expressed as a means for taking that action or as anelement which causes that action. Similarly, each physical elementdisclosed should be understood to encompass a disclosure of the actionwhich that physical element facilitates. Regarding this last aspect, asbut one example, the disclosure of a “dispenser” should be understood toencompass disclosure of the act of “dispensing”—whether explicitlydiscussed or not—and, conversely, were there effectively disclosure ofthe act of “dispensing”, such a disclosure should be understood toencompass disclosure of a “dispenser” and even a “means for dispensing.”Such changes and alternative terms are to be understood to be explicitlyincluded in the description.

Any patents, publications, or other references mentioned in thisapplication for patent are hereby incorporated by reference. Inaddition, as to each term used it should be understood that unless itsutilization in this application is inconsistent with a broadlysupporting interpretation, common dictionary definitions should beunderstood as incorporated for each term and all definitions,alternative terms, and synonyms such as contained in the Random HouseWebster's Unabridged Dictionary, second edition are hereby incorporatedby reference. Finally, all references listed in following list ofreferences or other documents or references filed with the applicationare hereby appended and hereby incorporated by reference, however, as toeach of these, to the extent that such information or statementsincorporated by reference might be considered inconsistent with thepatenting of this/these invention(s) such statements are expressly notto be considered as made by the applicant(s):

I. U.S. Patent Documents

DOCUMENT NO. Pages, Columns, Lines Where & KIND CODE (if PUB'N DATEPATENTEE OR Relevant Passages Or Relevant known) mm-dd-yyyy APPLICANTNAME Drawings Appear 2,039,219 04-28-1936 Hausser, et al. 2002/0142470A1 10-03-2002 Clarke et al. 2002/0182623 A1 12-05-2002 Lefevre et al.2003/0124729 A1 07-03-2003 Christensen et al. 2003/0138877 A1 07-24-2003Gibbs et al. 2003/0203493 A1 10-30-2003 Lemme et al. 2004/0033163 A102-19-2004 Tseung et al. 2004/0043495 A1 03-02-2004 Stokes et al.2004/0191128 A1 09-30-2004 Bogen et al. 2004/0241050 A1 12-02-2004 Bogenet al. 3,777,283 12-04-1973 Elkins 4,034,700 07-12-1977 Bassett et al.4,043,292 08-23-1977 Rogers et al. 4,199,613 04-22-1980 Johnson4,378,333 03-29-1983 Laipply 4,501,496 02-26-1985 Griffin 4,505,55703-19-1985 Golias 4,731,335 03-15-1988 Brigati 4,777,020 10-11-1988Brigati 4,790,640 12-13-1988 Nason 4,801,431 01-31-1989 Cuomo et al.4,847,208 07-11-1989 Bogen 4,985,206 01-15-1991 Bowman et al. 5,023,18706-11-1991 Koebler et al. 5,068,091 11-26-1991 Toya 5,338,358 08-16-1994Mizusawa et al. 5,425,918 06-20-1995 Healey et al. 5,569,607 10-29-1996Simon et al. 5,650,327 07-22-1977 Copeland et al. 5,654,199 08-05-1997Copeland et al. 5,654,200 08-05-1997 Copeland et al. 5,804,14109-08-1998 Chianese 5,839,091 11-17-1998 Rhett et al. 5,948,35809-07-1999 Kalra et al. 6,096,271 08-01-2000 Bogen et al. 6,165,73912-26-2000 Clatch 6,183,693 B1 02-06-2001 Bogen et al. 6,218,191 B104-17-2001 Palander 6,296,809 B1 10-02-2001 Richards et al. 6,349,264 B102-19-2002 Rhett et al. 6,352,861 B1 03-05-2002 Copeland et al.6,495,106 B1 12-17-2002 Kalra et al. 6,541,261 B1 04-01-2003 Bogen etal. 6,544,798 B1 04-08-2003 Christensen et al. 6,582,962 B1 06-24-2003Richards et al. 6,703,247 B1 03-09-2004 Chu 6,735,531 B2 05-11-2004Rhett et al. 6,735,531 B2 05-11-2004 Rhett et al. 6,746,851 B106-08-2004 Tseung et al. 6,783,733 B2 08-31-2004 Bogen et al, 6,827,901B2 12-07-2004 Copeland et al.II. Other DocumentsU.S. Provisional Application No. 60/673,468 filed Apr. 21, 2005,Entitled Method and Apparatus for Automated Rapid Immunohistochemistry

Thus, the applicant should be understood to have support to claim andmake a statement of invention to at least: i) each of the processingdevices as herein disclosed and described, ii) the related methodsdisclosed and described, iii) similar, equivalent, and even implicitvariations of each of these devices and methods, iv) those alternativedesigns which accomplish each of the functions shown as are disclosedand described, v) those alternative designs and methods which accomplisheach of the functions shown as are implicit to accomplish that which isdisclosed and described, vi) each feature, component, and step shown asseparate and independent inventions, vii) the applications enhanced bythe various systems or components disclosed, viii) the resultingproducts produced by such systems or components, ix) each system,method, and element shown or described as now applied to any specificfield or devices mentioned, x) methods and apparatuses substantially asdescribed hereinbefore and with reference to any of the accompanyingexamples, xi) the various combinations and permutations of each of theelements disclosed, and xii) each potentially dependent claim or conceptas a dependency on each and every one of the independent claims orconcepts presented. In addition and as to computer aspects and eachaspect amenable to programming or other electronic automation, theapplicant should be understood to have support to claim and make astatement of invention to at least: xiii) processes performed with theaid of or on a computer as described throughout the above discussion,xiv) a programmable apparatus as described throughout the abovediscussion, xv) a computer readable memory encoded with data to direct acomputer comprising means or elements which function as describedthroughout the above discussion, xvi) a computer configured as hereindisclosed and described, xvii) individual or combined subroutines andprograms as herein disclosed and described, xviii) the related methodsdisclosed and described, xix) similar, equivalent, and even implicitvariations of each of these systems and methods, xx) those alternativedesigns which accomplish each of the functions shown as are disclosedand described, xxi) those alternative designs and methods whichaccomplish each of the functions shown as are implicit to accomplishthat which is disclosed and described, xxii) each feature, component,and step shown as separate and independent inventions, and xxiii) thevarious combinations and permutations of each of the above.

With regard to claims now or later presented for examination, it shouldbe understood that for practical reasons and so as to avoid greatexpansion of the examination burden, the applicant may at any timepresent only initial claims or perhaps only initial claims with onlyinitial dependencies. Support should be understood to exist to thedegree required under new matter laws—including but not limited toEuropean Patent Convention Article 123(2) and United States Patent Law35 USC 132 or other such laws—to permit the addition of any of thevarious dependencies or other elements presented under one independentclaim or concept as dependencies or elements under any other independentclaim or concept. In drafting any claims at any time whether in thisapplication or in any subsequent application, it should also beunderstood that the applicant has intended to capture as full and broada scope of coverage as legally available. To the extent thatinsubstantial substitutes are made, to the extent that the applicant didnot in fact draft any claim so as to literally encompass any particularembodiment, and to the extent otherwise applicable, the applicant shouldnot be understood to have in any way intended to or actuallyrelinquished such coverage as the applicant simply may not have beenable to anticipate all eventualities; one skilled in the art, should notbe reasonably expected to have drafted a claim that would have literallyencompassed such alternative embodiments.

Further, if or when used, the use of the transitional phrase“comprising” is used to maintain the “open-end” claims herein, accordingto traditional claim interpretation. Thus, unless the context requiresotherwise, it should be understood that the term “comprise” orvariations such as “comprises” or “comprising”, are intended to implythe inclusion of a stated element or step or group of elements or stepsbut not the exclusion of any other element or step or group of elementsor steps. Such terms should be interpreted in their most expansive formso as to afford the applicant the broadest coverage legally permissible.

Finally, any claims set forth at any time are hereby incorporated byreference as part of this description of the invention, and theapplicant expressly reserves the right to use all of or a portion ofsuch incorporated content of such claims as additional description tosupport any of or all of the claims or any element or component thereof,and the applicant further expressly reserves the right to move anyportion of or all of the incorporated content of such claims or anyelement or component thereof from the description into the claims orvice-versa as necessary to define the matter for which protection issought by this application or by any subsequent continuation, division,or continuation-in-part application thereof, or to obtain any benefitof, reduction in fees pursuant to, or to comply with the patent laws,rules, or regulations of any country or treaty, and such contentincorporated by reference shall survive during the entire pendency ofthis application including any subsequent continuation, division, orcontinuation-in-part application thereof or any reissue or extensionthereon.

1. A method of biochemical processing comprising the steps of: a. obtaining at least one sample; b. selecting a biochemical test sequence for said at least one sample; c. establishing said at least one sample on a proximally paired sample holder; d. snapping in at least one source in a sample processing system; e. establishing at least one consumable biochemical process element consumably responsive to said biochemical test sequence; f. establishing at least one consumable biochemical process alterable information memory element associated with said consumable biochemical process element; g. detachably electrically connecting said consumable biochemical process alterable information memory element to said sample processing system; h. querying said consumable biochemical process alterable information memory element; i. subjecting at least a portion of an exterior sample area of said at least one sample to an appropriate fluidic substance for said biochemical test sequence; j. establishing a firmly bounded fluidic environment in the vicinity of said exterior sample area at least in part through the presence of said fluidic substance; k. causing hinged movement between a first surface relative to a second surface; l. automatically affirmatively initiating a fluid wave in said firmly bounded fluidic environment as a result of said hinged movement between a first surface relative to a second surface; m. automatically substantially stopping said fluid wave in said firmly bounded fluidic environment; n. automatically moving an absorbent material to a position in the vicinity of said sample holder; o. contacting said absorbent material and said fluidic substance; p. automatically withdrawing said fluidic substance from proximity to said sample upon completion of at least a portion of said process; q. automatically processing said biochemical test sequence; r. changing at least some information on said consumable biochemical process alterable information memory element as a result of actions conducted in said step of automatically processing said biochemical test sequence; and s. accomplishing desired results through said biochemical test sequence.
 2. A method of biochemical processing as described in claim 1 wherein said step of snapping in at least one source comprises the step of snapping in at least one source having substances selected from a group consisting of: a primary antibody substance, a secondary antibody substance, a chromogen substance, a counterstain substance, and a buffer substance.
 3. A method of biochemical processing as described in claim 2 wherein said step of snapping in at least one source comprises the step of snapping in at least two sources.
 4. A method of biochemical processing as described in claim 3 wherein said step of snapping in at least two sources comprises the step of: a. snapping in a primary antibody substance cartridge; and b. snapping in a linear multiple reagent magazine.
 5. A method of biochemical processing as described in claim 1 wherein said step of snapping in at least one source comprises the step of utilizing at least one location specific relative substance use sized source.
 6. A method of biochemical processing as described in claim 1, further comprising the step of interfacing with a laboratory information system.
 7. A method of biochemical processing comprising the steps of: a. obtaining at least one sample; b. selecting a biochemical test sequence for said at least one sample; c. establishing at least one substance withdrawal consumable biochemical process element consumably responsive to said biochemical test sequence; d. establishing at least one consumable biochemical process alterable information memory element associated with said substance withdrawal consumable biochemical process element; e. detachably electrically connecting said consumable biochemical process alterable information memory element to a sample processing system; f. querying said consumable biochemical process alterable information memory element: g. subjecting at least a portion of an exterior sample area of said at least one sample to an appropriate substance for said biochemical test sequence; h. automatically processing said biochemical test sequence; i. changing at least some information on said consumable biochemical process alterable information memory element as a result of actions conducted in said step of automatically processing said biochemical test sequence; and j. accomplishing desired results through said biochemical test sequence
 8. A method of biochemical processing as described in claim 7 wherein said step of establishing at least one substance withdrawal consumable biochemical process element comprises the step of encasing an absorbent material in a confinement enclosure.
 9. A method of biochemical processing as described in claim 8 and further comprising the step of automatically sequencing said absorbent material to establish an unused portion of absorbent material in an exposed position as part of said biochemical test sequence. 